The present invention relates generally to imaging of objects. In particular, the present invention provides methods and systems for imaging one or more objects using one or more pulses of particles containing from about one electron to about 10,000 electrons or more preferably about 10 electrons to about 100 electrons in a transmission electron microscope system. More particularly, the present invention provides methods and systems for identifying information about one or more temporal components associated with one or more spatial features of certain objects being imaged. Merely by way of example, the invention has been applied to imaging certain chemical, physical, and biological objects. The invention, however, can also be applied to other applications such as other areas of biology, chemistry (e.g., organic, physical, biochemistry), medicine (e.g., medical devices, diagnostics, analysis, treatments), physical sciences, electronics, semiconductor devices and materials (e.g., silicon, germanium, Group III/V, Group II/VI), chemicals (e.g., industrial), petrochemical (e.g., gas, oil), any combination of these, and the like. The invention may be applied to applications involving processing and/or screening of certain compounds and/or molecules such as an oligomer, a peptide, a nucleic acid, an oligosaccharide, a phospholipid, a polymer, a protein, or a drug congener preparation, or any other like species and/or entities. Additionally, the invention may be applied to diffraction, crystallography, spectroscopy, and other areas, and the like. Still further, the invention may be applied to monitoring formation/decomposition of materials, film(s), compounds, and/or other species, depending upon the embodiment.
The twentieth century has been witness to certain major advances in our ability to peer into the microscopic world of molecules, thereby giving us unparalleled insights into their static behavior. For example, electron microscopes, particularly the transmission electron microscope (TEM), provide for direct imaging of macromolecular static structures with spatial resolution of a few angstroms. These conventional electron microscope systems have been employed to image, for example, biological macromolecular crystals. Despite these advances in imaging, the dynamic aspects of structural evolution, insight into which is desirable in understanding function, cannot be obtained from currently available static images because of the lack of temporal resolution in conventional electron microscopy.
When chemical, and especially biological changes, involve complex transient structures with many possible conformations, one must often address the nature of the three-dimensional (3D) molecular structures, but at different times during the change. In most biologically important processes, the structural change is reversible, which means that upon initiation of an event and the associated response, a nondestructive reaction occurs that is repeatable. Unfortunately, limitations exist with conventional microscopic techniques. As an example, conventional electron microscopes cannot generally produce images with spatial resolution on the biological length scale (ranging from nanometers to micrometers) with desirable temporal resolutions. These and other limitations of conventional techniques are described throughout the present specification and more particularly below.
Thus, there is a need in the art for improved methods imaging at atomic scale resolutions for physical, chemical, biological, and other samples.